Disposition of citalopram in biological specimens from postmortem cases.
Citalopram is a bicyclic phthalate compound approved in 1998 by the U.S. Food and Drug Administration for the treatment of depression. It is a highly selective serotonin reuptake inhibitor that appears to have little effect on noradrenaline or dopamine reuptake. Since this drug has only recently been released on the U.S. market, information regarding therapeutic, toxic, and lethal concentrations is sparse. This study reports the detection of citalopram in 22 postmortem cases. Citalopram was identified and quantitated by capillary column gas chromatography with nitrogen phosphorus detection after basic liquid-liquid extraction. Confirmation was achieved by full scan electron impact gas chromatography/mass spectrometry. In the 22 cases studied, heart blood citalopram concentrations ranged from 0.09 to 1.64 mg/L (n = 22, mean +/- SD = 0.51+/-0.43, median = 0.34); femoral blood concentrations ranged from 0.09 to 0.76 mg/L (n = 14, mean +/- SD = 0.34+/-0.23, median = 0.28); and urine concentrations ranged from 0.05 to 276.00 mg/L (n = 13). Liver was analyzed in three cases with citalopram concentrations ranging from 2.22 to 8.08 mg/kg. The average heart blood/femoral blood ratio was 1.26 (range 0.75 to 1.98, n = 14). In each case, the cause of death was not considered to be related to citalopram toxicity. These data may therefore provide a basis for establishing post mortem citalopram concentrations following therapeutic doses.


Monodisperse Long-Chain Sulfobetaine-Capped CsPbBr3 Nanocrystals and Their Superfluorescent Assemblies.
Ligand-capped nanocrystals (NCs) of lead halide perovskites, foremost fully inorganic CsPbX3 NCs, are the latest generation of colloidal semiconductor quantum dots. They offer a set of compelling characteristics-large absorption cross section, as well as narrow, fast, and efficient photoluminescence with long exciton coherence times-rendering them attractive for applications in light-emitting devices and quantum optics. Monodisperse and shape-uniform, broadly size-tunable, scalable, and robust NC samples are paramount for unveiling their basic photophysics, as well as for putting them into use. Thus far, no synthesis method fulfilling all these requirements has been reported. For instance, long-chain zwitterionic ligands impart the most durable surface coating, but at the expense of reduced size uniformity of the as-synthesized colloid. In this work, we demonstrate that size-selective precipitation of CsPbBr3 NCs coated with a long-chain sulfobetaine ligand, namely, 3-(N,N-dimethyloctadecylammonio)-propanesulfonate, yields monodisperse and sizable fractions (>100 mg inorganic mass) with the mean NC size adjustable in the range between 3.5 and 16 nm and emission peak wavelength between 479 and 518 nm. We find that all NCs exhibit an oblate cuboidal shape with the aspect ratio of 1.2 × 1.2 × 1. We present a theoretical model (effective mass/k·p) that accounts for the anisotropic NC shape and describes the size dependence of the first and second excitonic transition in absorption spectra and explains room-temperature exciton lifetimes. We also show that uniform zwitterion-capped NCs readily form long-range ordered superlattices upon solvent evaporation. In comparison to more conventional ligand systems (oleic acid and oleylamine), supercrystals of zwitterion-capped NCs exhibit larger domain sizes and lower mosaicity. Both kinds of supercrystals exhibit superfluorescence at cryogenic temperatures-accelerated collective emission arising from the coherent coupling of the emitting dipoles.


Activation of Akt characterizes estrogen receptor positive human breast cancers which respond to anthracyclines.
Anthracyclines are key components of human breast cancer chemotherapy. Here, we explored the role of Akt signaling in anthracycline resistance.The antitumor activity of doxorubicin and Akt inhibitor A-443654 alone or combined was examined in estrogen receptor (ER) positive and negative human breast cancer cell lines. Further, we examined mRNA changes induced by anthracyclines in locally advanced breast cancers biopsied before and after treatment in two clinical trials.Doxorubicin increased Akt phosphorylation in ER positive MCF7 and T47D cell lines, with no effect in ER negative MDA-MB231 breast cancer cells. A-443654 was significantly more cytotoxic in doxorubicin-resistant compared to doxorubicin-naïve MCF7. This difference was not observed in MDA-MB231. Among 24 patients, AKT1 gene expression increased 24 hrs after the initial epirubicin exposure in ER positive tumors responding to therapy (n=6), as compared to ER positive non-responders (n=7) or ER negative tumors (n=11). In contrast, AKT1 mRNA changes after 16 weeks of doxorubicin were unrelated to clinical response and ER status (n=30).In conclusion, rapid Akt activation was observed in ER positive breast cancers which responded to anthracyclines. Increased cytotoxicity of A-443654 in doxorubicin-resistant MCF7 cells indicates a possible role for Akt inhibitors in ER positive breast cancers where chemoresistance evolves.


Clinical outcome and imaging changes after intraarticular (IA) application of etanercept or methylprednisolone in rheumatoid arthritis: magnetic resonance imaging and ultrasound-Doppler show no effect of IA injections in the wrist after 4 weeks.
To assess the magnetic resonance imaging (MRI) and ultrasound (US) changes in the wrist of patients with rheumatoid arthritis (RA) 4 weeks after an US guided intraarticular (IA) injection. Contrast enhanced MRI and US-Doppler were performed at baseline and 4 weeks after IA injection of either 40 mg methylprednisolone (n = 12) or 25 mg etanercept (n = 13) in 25 patients with RA taking disease modifying antirheumatic drugs with a therapy-resistant wrist joint. All injections were US guided. There was an improvement in swollen target joint score (p < 0.001), tender target joint score (p < 0.002), and physician visual analog scale score (p < 0.001) after 4 weeks. Baseline MRI synovitis score was mean 5.08 (range 3-9) and was unchanged at followup in the whole group (p = 0.52) and between treatment groups (p = 0.43). MRI edema score (mean 4.46, range 0-29) in the total group was unchanged after 4 weeks (p = 0.13), whereas MRI erosion score increased in the total group from baseline, 17.88 (range 7-40), to 4 weeks, 18.25 (range 7-40) (p < 0.001). Neither US-Doppler color fraction (0.07) nor Resistive Index (RI) (p = 0.36) changed from baseline to 4 week followup. In contrast to the clinical evaluation, imaging measures of relevance for the estimation of inflammation, US-Doppler, US RI, MRI synovitis, and bone-marrow edema did not change 4 weeks after a single IA injection of either methylprednisolone or etanercept in the wrist. Within the same period, erosive progression in some patients suggested that joints with active disease may deteriorate within as little as 1 month, and that this development is not arrested by 1 injection. Given the small sample size of our study further studies are required to confirm our results.


A combined NMR and computational approach to investigate peptide binding to a designed Armadillo repeat protein.
The specific recognition of peptide sequences by proteins plays an important role both in biology and in diagnostic applications. Here we characterize the relatively weak binding of the peptide neurotensin (NT) to the previously developed Armadillo repeat protein VG_328 by a multidisciplinary approach based on solution NMR spectroscopy, mutational studies, and molecular dynamics (MD) simulations, totaling 20μs for all MD runs. We describe assignment challenges arising from the repetitive nature of the protein sequence, and we present novel approaches to address them. Partial assignments obtained for VG_328 in combination with chemical shift perturbations allowed us to identify the repeats not involved in binding. Their subsequent elimination resulted in a reduced-size binder with very similar affinity for NT, for which near-complete backbone assignments were achieved. A binding mode suggested by automatic docking and further validated by explicit solvent MD simulations is consistent with paramagnetic relaxation enhancement data collected using spin-labeled NT. Favorable intermolecular interactions are observed in the MD simulations for the residues that were previously shown to contribute to binding in an Ala scan of NT. We further characterized the role of residues within the N-cap for protein stability and peptide binding. Our multidisciplinary approach demonstrates that an initial low-resolution picture for a low-micromolar-peptide binder can be refined through the combination of NMR, protein design, docking, and MD simulations to establish its binding mode, even in the absence of crystallographic data, thereby providing valuable information for further design.


Improvement in functional vocabulary and generalization to conversation following a self-administered treatment using a smart tablet in primary progressive aphasia.
Currently, public services in speech-language pathology for primary progressive aphasia (PPA) are very limited, although several interventions have been shown to be effective. In this context, new technologies have the potential to enable people with PPA to improve their communication skills. The main aim of this study was to investigate the efficacy of a self-administered therapy using a smart tablet to improve naming of functional words and to assess generalization to an ecological conversation task. Five adults with PPA completed the protocol. Using an ABA design with multiple baselines, naming performance was compared across four equivalent lists: (1) trained with functional words; (2) trained with words from a picture database; (3) exposed but not trained; and (4) not exposed (control). Treatment was self-administered four times a week for a period of four consecutive weeks. A significant improvement for trained words was found in all five participants, and gains were maintained two months post-treatment in four of them. Moreover, in three participants, evidence of generalization was found in conversation. This study supports the efficacy of using a smart tablet to improve naming in PPA and suggests the possibility of generalization to an ecological context.


Establishment of a Novel Scoring System for Colon Capsule Endoscopy to Assess the Severity of Ulcerative Colitis-Capsule Scoring of Ulcerative Colitis.
The usefulness of second-generation colon capsule endoscopy (CCE-2) for ulcerative colitis (UC) has not been fully demonstrated. This study aimed to develop an endoscopic severity score of UC for CCE-2. Patients diagnosed with UC were enrolled prospectively and underwent colonoscopy and CCE-2 on the same day. The collected CCE-2 videos were adopted for the development of the score. These videos were scored by 4 blinded inflammatory bowel disease experts. The items validated with the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) were used as the candidate items, some of which were automatically assessed using the workstation. Each item was divided into proximal and distal parts at the splenic flexure and then individually assessed. The image readers simultaneously evaluated the inflammation severity using the visual analog scale (VAS). The descriptors that contribute to this scale were evaluated, and a model to predict the VAS was constructed. The UCEIS was scored by other endoscopists using colonoscopy videos. The correlation coefficients with fecal calprotectin, blood tests, and Lichtiger index were calculated. The final scoring system was fixed as "vascular pattern sum (proximal + distal) + bleeding sum + erosions and ulcers sum (minimum-maximum, 0-14)" and was named Capsule Scoring of Ulcerative Colitis (CSUC). The correlation coefficient of CSUC with biomarkers and clinical score was similar to that of the UCEIS. We developed a new simple score using the 3 descriptors of CCE-2.


Mitochondrial DNA in the oocyte and the developing embryo.
Mitochondria play a primary role in cellular energetic metabolism, homeostasis, and death. They possess their own multicopy genome, which is maternally transmitted. Mitochondria are directly involved at several levels in the reproductive process since their functional status influences the quality of oocytes and contributes to the process of fertilization and embryonic development. This chapter discusses recent findings concerning mitochondrial DNA content and its expression during oogenesis, fertilization, and early embryonic development.


Successful radiation treatment of undertail fibrosarcoma in a Major Mitchell's cockatoo (Lophochroa leadbeateri).
A male Major Mitchell's cockatoo (Lophochroa leadbeateri) of unknown age presented with an ulcerated mass on the ventral tail caudodorsal to the cloaca. An impression smear of the mass showed spindle cell atypia. Multiple biopsies were submitted for histopathology with inconclusive results. A CT scan revealed a soft tissue mass causing compression of the cloacal lumen. The patient underwent surgical debulking, and a core of the mass was submitted again for histopathology, which reported it as fibrosarcoma. Under repeated general isoflurane gas anesthesia, the patient received a course of definitive radiation therapy totaling 60 Gy and divided in 3 Gy X 20 fractions. By treatment completion, the lesion had decreased in size with necrotic debris on the surface. Surrounding tissues appeared healthy and no adverse effects were observed. As of 1.5 years post-treatment, the mass appears completely healed with no signs of reoccurrence. This case suggests that radiation therapy with this protocol could be an effective treatment option for fibrosarcoma in avian species.


Quality issues concerning genetic counselling for presymptomatic testing: a European Delphi study.
Genetic counselling for presymptomatic testing is complex, bringing both ethical and practical questions. There are protocols for counselling but a scarcity of literature regarding quality assessment of such counselling practice. Generic quality assessment tools for genetic services are not specific to presymptomatic testing (PST). Therefore, the aim of this study was to identify aspects of effective counselling practice in PST for late-onset neurological disorders. We used the Delphi method to ascertain the views of relevant European experts in genetic counselling practice, ascertained via published literature and nomination by practitioners. Ethical approval was obtained. Questionnaires were sent electronically to a list of 45 experts, (Medical Doctors, Geneticists, Genetic Counsellors and Genetic Nurses), who each contributed to one to three rounds. In the first round, we provided a list of relevant indicators of quality of practice from a literature review. Experts were requested to evaluate topics in four domains: (a) professional standards; (b) service standards; (c) the consultant's perspective; and (d) protocol standards. We then removed items receiving less than 65% approval and added new issues suggested by experts. The second round was performed for the refinement of issues and the last round was aimed at achieving final consensus on high-standard indicators of quality, for inclusion in the assessment tool. The most relevant indicators were related to (1) consultant-centred practice and (2) advanced counselling and interpersonal skills of professionals. Defined high-standard indicators can be used for the development of a new tool for quality assessment of PST counselling practice.


Efficacy of dry needling as an adjunct to manual therapy for patients with chronic mechanical neck pain: a randomised clinical trial.
Chronic mechanical neck pain is associated with musculoskeletal tissue alterations. Active trigger points in the trapezius and levator scapulae muscles are common in patients with chronic mechanical neck pain. In this study, we compared the effect of dry needling (DN) combined with manual therapy (MT) to sham dry needling (SDN) combined with MT on pain, pain pressure threshold, cervical range of motion and neck disability in patients with chronic mechanical neck pain. A randomised, single-blind clinical trial was carried out involving 101 participants with chronic mechanical neck pain, divided into an intervention group (DN+MT, n=47) and a control group (SDN+MT, n=54). Participants received two treatment sessions. The intervention group received MT in conjunction with DN of the most mechano-sensitive myofascial trigger point (MTrP). The control group received MT plus SDN. Outcomes measures were: pain intensity (numeric pain rating scale, NPRS), pressure pain threshold (PPT), cervical range of motion (ROM) and neck disability (neck disability index, NDI). This study found that between-group differences in pain intensity were statistically significant (P<0.01). Pain decreased after the first intervention in the DN+MT group (3.47±0.25 points on the NPRS) and even more so after the second intervention (4.76±0.24 points on the NPRS). After 4 weeks, pain intensity differed from baseline by 4.89±0.27 points on the NPRS. Statistically significant differences (P<0.001) in PPT were also found between the intervention group and the control group. After the first intervention, a significant increase in PPT within the DN+MT group (3.09±0.8 kg/cm2) was observed. Cervical ROM also showed highly statistically significant differences. After 4 weeks, a statistically significant reduction (P<0.001) in NDI was observed between the two groups. Our results show that DN+MT is efficacious and significantly better than SDN+MT at reducing pain intensity, PPT, neck disability and cervical ROM in patients with chronic mechanical neck pain. 1b.


Effect of experimental diabetes on the susceptibility of myelin proteins to proteolysis.
To investigate abnormalities in peripheral nerve myelin in experimental diabetes, we studied the effects of the proteolytic enzymes trypsin and alpha-chymotrypsin on the proteins of this membrane, obtained from the sciatic nerves of normal rats and from animals made diabetic with streptozotocin. The dominant effect of the proteolytic enzymes was to incompletely degrade the major membrane protein (mol. wt. 28,000), with the appearance of new protein (mol. wt. 20,000). Using myelin isolated from the nerves of diabetic animals, the reaction was approximately one-half that of the controls (P less than 0.01) for both enzymes. When, however, the myelin protein affected by trypsin and chymotrypsin was isolated from the membrane and then incubated with the proteolytic enzyme, its proteolysis was complete and took place at the same rate in the diabetic animals and controls. These findings suggest that, in this model of experimental diabetes, there is an alteration in the structure of peripheral nerve myelin that inhibits interaction between the protein in the membrane bilayer and two water soluble proteolytic enzymes. This alteration could not be demonstrated in protein isolated from the membrane, suggesting that the change relates to the interaction of the protein and other components of myelin, rather than to chemical alteration in the protein per se.


Diversification in the HIV-1 Envelope Hyper-variable Domains V2, V4, and V5 and Higher Probability of Transmitted/Founder Envelope Glycosylation Favor the Development of Heterologous Neutralization Breadth.
A recent study of plasma neutralization breadth in HIV-1 infected individuals at nine International AIDS Vaccine Initiative (IAVI) sites reported that viral load, HLA-A*03 genotype, and subtype C infection were strongly associated with the development of neutralization breadth. Here, we refine the findings of that study by analyzing the impact of the transmitted/founder (T/F) envelope (Env), early Env diversification, and autologous neutralization on the development of plasma neutralization breadth in 21 participants identified during recent infection at two of those sites: Kigali, Rwanda (n = 9) and Lusaka, Zambia (n = 12). Single-genome analysis of full-length T/F Env sequences revealed that all 21 individuals were infected with a highly homogeneous population of viral variants, which were categorized as subtype C (n = 12), A1 (n = 7), or recombinant AC (n = 2). An extensive amino acid sequence-based analysis of variable loop lengths and glycosylation patterns in the T/F Envs revealed that a lower ratio of NXS to NXT-encoded glycan motifs correlated with neutralization breadth. Further analysis comparing amino acid sequence changes, insertions/deletions, and glycan motif alterations between the T/F Env and autologous early Env variants revealed that extensive diversification focused in the V2, V4, and V5 regions of gp120, accompanied by contemporaneous viral escape, significantly favored the development of breadth. These results suggest that more efficient glycosylation of subtype A and C T/F Envs through fewer NXS-encoded glycan sites is more likely to elicit antibodies that can transition from autologous to heterologous neutralizing activity following exposure to gp120 diversification. This initiates an Env-antibody co-evolution cycle that increases neutralization breadth, and is further augmented over time by additional viral and host factors. These findings suggest that understanding how variation in the efficiency of site-specific glycosylation influences neutralizing antibody elicitation and targeting could advance the design of immunogens aimed at inducing antibodies that can transition from autologous to heterologous neutralizing activity.


The polyphenol 3, 4, 5 - tri-hydroxy benzoic acid inhibits indian daboia russelli venom and its hemorrhagic complex induced local toxicity.
Despite a long history on treatment and management of snakebite, as of now, no satisfactory cure exists to treat local toxicity, including anti-venom therapy. Several natural compounds from plants and their synthetic analogs have shown to be protective. In this study 3, 4, 5-tri-hydroxy benzoic acid, the gallic acid (GA) was tested against the local toxicity of Daboia russelli (DR) venom and its purified hemorrhagic complex (HC). GA inhibited in vitro proteolytic activity of both DR venom and HC but, it did not inhibit phospholipase activity of DR venom. GA inhibited hemorrhage, edema forming, dermo- and myonecrotic activities of both HC and DR venom in in vivo experiments. GA was particularly effective against hemorrhagic activity but, GA inhibition had a greater effect on HC when compared to DR venom. The inhibition was likely due to GA induced structural changes in HC as revealed by alterations in fluorescence emission and CD spectral properties. However, the inhibition was not due to chelating property of GA as suggested by UV-visible spectral studies. Inhibition of collagen type IV, laminin and fibronectin degradation essentially provided the biochemical basis for GA which inhibited local effects of HC as well as DR venom. Thus, the study appears highly promising to explore GA and its generics against ruthless local effects and perhaps systemic hemorrhage of DR and other snake bites as well. Further, these agents will possibly find an immense value in the regulation of matrix metalloproteases (MMPs) in processes such as wound healing, inflammation and in the treatment of cancer.


Diagnostic consistency and interchangeability of schizophrenic disorders and bipolar disorders: A 7-year follow-up study.
The change in psychiatric diagnoses in clinical practice is not an unusual phenomenon. The interchange between the diagnoses of schizophrenic disorders and bipolar disorders is a major clinical issue because of the differences in treatment regimens and long-term prognoses. In this study, we used a nationwide population-based sample to compare the diagnostic consistency and interchange rate between schizophrenic disorders and bipolar disorders. In total, 25 711 and 11 261 patients newly diagnosed as having schizophrenic disorder and bipolar disorder, respectively, were retrospectively enrolled from the Psychiatric Inpatient Medical Claims database between 2001 and 2005. We followed these two cohorts for 7 years to determine whether their diagnoses were consistent throughout subsequent hospitalizations. The interchange between the two diagnoses was analyzed. In the schizophrenic disorder cohort, the overall diagnostic consistency rate was 87.3% and the rate of change to bipolar disorder was 3.0% during the 7-year follow-up. Additional analyses of subtypes revealed that the change rate from schizoaffective disorder to bipolar disorder was 12.0%. In the bipolar disorder cohort, the overall diagnostic consistency rate was 71.9% and the rate of change to schizophrenic disorder was 8.3%. Changes in the diagnosis of a major psychosis are not uncommon. The interchange between the diagnoses of schizophrenic disorders and bipolar disorders might be attributed to the evolution of clinical symptoms and the observation of preserved social functions that contradict the original diagnosis. While making a psychotic diagnosis, clinicians should be aware of the possibility of the change in diagnosis in the future.


Effect of Interpregnancy Interval on Adverse Perinatal Outcomes in Southern China: A Retrospective Cohort Study, 2000-2015.
In January 2016, a universal two-child policy was introduced in China. The association of interpregnancy interval (IPI) with perinatal outcomes has not previously been assessed among Chinese population. We investigated the effect of IPI after live birth on the risks of preterm delivery, and small, and large for gestational age births in China. We conducted a cohort study among 227 352 Chinese women with their first and second delivery during 2000 to 2015. IPI was calculated as months from first live delivery to conception of the second pregnancy. Poisson regression models with robust variance were fit to evaluate associations of IPI with risk of adverse perinatal outcomes, adjusted for potential confounders. Compared to IPI of 24- <30 months, IPI <18 months was associated with higher risks of preterm birth (PTB) and small for gestational age (SGA). For IPI <6 months, the adjusted relative risks (RR) for PTB and SGA were 2.04 (95% confidence interval [CI] 1.83, 2.27) and 1.43 (95% CI 1.31, 1.57), respectively. Women with IPI ≥60 months had higher risks of PTB and large for gestational age (LGA). For IPI ≥120 months, the adjusted RRs for PTB and LGA were 1.67 (95% CI 1.43, 1.94) and 1.10 (95% CI 0.97, 1.26). Women with IPI <18 months after live birth had higher risk of PTB and SGA, and IPI ≥60 months was associated with higher risk of PTB and LGA. These findings may provide information to Chinese couples about the appropriate interpregnancy interval for a second pregnancy.


Are rich people or poor people more likely to be ill? Lay perceptions, by social class and neighbourhood, of inequalities in health.
Research in the UK has suggested that people in lower social classes or from poorer neighbourhoods are less likely than their more socially advantaged counterparts to agree that health and life expectancy are worse among more deprived population groups. The small body of previous research has either used qualitative approaches or coded open-ended responses to survey questions about causes of health and illness or of inequalities between areas. We examined lay perceptions by asking a direct question and using a quantitative, multivariate approach. Residents in three age groups (25, 45 and 65 years old) living in two socially contrasting localities in Glasgow, Scotland, were asked who were more likely to have accidents, cancer, heart disease, mental illness, to be fitter, and to live longer: rich people, poor people, or both equally. Across all the health categories, those in lower social classes or from poorer neighbourhoods were equally or less likely than their more socially advantaged counterparts to say the poor had worse health. In a model containing age, sex, class and locality, those in lower social classes and in the poorer locality were significantly less likely to say that richer people live longer (OR: 0.5). We have therefore confirmed earlier observations that those most at risk of ill health may be less likely to acknowledge the social gradient in health. We suggest a need to examine this apparent paradox in other contexts and in more detail, using both quantitative and qualitative approaches.


Accessing osteocyte lacunar geometrical properties in human jaw bone on the submicron length scale using synchrotron radiation μCT.
The architectural properties of the osteocyte cell network provide a valuable basis for understanding the mechanisms of bone remodelling, mineral homeostasis, ageing and pathologies. Recent advances in synchrotron microtomography enable unprecedented three-dimensional imaging of both the bone lacunar network and the extracellular matrix. Here, we investigate the three-dimensional morphological properties of osteocyte lacunae in human healthy and bisphosphonate-related osteonecrotic jaw bone based on synchrotron X-ray computed tomography images, with a spatial isotropic voxel size of 300 nm. Bisphosphonate-related osteonecrosis of the jaw is a relatively new disease with increasing incidence, which remains poorly understood. A step forward in elucidating this malady is to assess whether, and how, the morphology of the osteocyte lacunar network is modified in the affected jaw tissue. We evaluate thousands of cell lacunae from five specimens of which three originate from patients diagnosed with bisphosphonate-associated osteonecrosis. In this exploratory study, we report three-dimensional quantitative results on lacunar volumes (296-502 μm(3)), shape (approximated by an ellipsoidal shape with principal axes a > b > c, such that a = 2.2b and a = 4c) and spatial distribution (i.e., 50% of the mineralized matrix volume is located within 12 μm to the closest lacunar boundary) at submicron resolution on such specimens. We observe that the average lacunar volumes of the bisphosphonate-related osteonecrotic jaw specimens were within the range of volumes found in the two specimens originating from healthy donors and conclude that lacunar volumes are not the key element in the course of bisphosphonate-related osteonecrotic jaw. In three out of five specimens we observe lacunar volume sizes in segmented osteons to be significantly different compared to lacunar volumes in the adjacent tissue regions. Furthermore, we quantify the number of lacunae containing small dense objects (on average 9%). In contrast to lacunar morphology we report the lacunar density (16,000-50,000 per mm(3)) to be different in jaw bone tissue compared to what has been reported in femoral sites.


Opening of a Peierls gap in BaVS3 probed by V L3 edge resonant inelastic x-ray scattering.
V L3 edge resonant inelastic x-ray scattering measurements performed on high quality BaVS3 single crystals reveal that the intra-t2g dd excitations close to the elastic peak are suppressed below the metal-insulator transition induced by the Peierls instability. The depletion of electronic states close to the Fermi level represents a direct observation of the opening of a charge gap inside the t2g manifold.


Modulating the pH-activity profile of cellulase A from Cellulomonas fimi by replacement of surface residues.
One industrial process for the production of cellulosic ethanol and or value-added products involves exposing the cellulose content of plant materials by steam explosion in the presence of strong acid, followed by its neutralization and subsequent digestion with a cocktail of cellulolytic enzymes. These enzymes typically have activity optima at slightly acidic or neutral pH and so generating enzymes that are more active and tolerant in more acidic conditions would help to reduce associated costs. Here, we describe the engineering of cellulase A from Cellulomonas fimi as a model to replace residues that were identified as potentially influencing the pH-activity profile of the enzyme based on sequence alignments and analysis of the known three-dimensional structures of other CAZy family 6 glycoside hydrolases with the aim to lower its pH optimum. Twelve specific residues and a sequence of eight were identified and a total of 30 mutant enzymes were generated. In addition to being replaced with natural amino acids, some of the identified residues were substituted with cysteine and subsequently oxidized to cysteinesulfinate. Of the four single amino acid replacements that produced enhancements of activity at acidic pH, three involved the removal of charged groups from the surface of the enzyme. The generation of double mutations provided mixed results but the combination of Glu407 → Ala and Tyr321 → Phe replacements had an additive effect on the enhancement, reaching a total activity that was 162% of the wild-type level. This study thus illustrated the utility of altering the surface charge properties of the family 6 glycoside hydrolases to enhance activity at low pH and thereby an avenue for further protein engineering.


Fumonisin B1 from the fungus Fusarium moniliforme causes contact toxicity in plants: evidence from studies with biosynthetically labeled toxin.
Fumonisin B1 (FB1) is the most abundant of a series of sphingosine analog mycotoxins produced by the fungus Fusarium moniliforme, a ubiquitous contaminant of stored corn (maize) worldwide. FB1 exhibits a variety of biological activities including phytotoxicity, which is of particular interest for its potential role as a virulence factor to facilitate invasion of plant tissues by the fungus. Droplets of FB1 solution applied to the leaf surface of jimsonweed, black nightshade, and susceptible tomatoes caused necrosis, growth inhibition, and death. With Arabidopsis thaliana grown on agar plates, an IC50 (concentration causing half maximal phytotoxicity) of less than 1 ppm was observed. [3H]FB1 was prepared by biosynthetic incorporation of commercially-available radiolabeled presumptive precursors into the toxin in rice medium solid cultures of F. moniliforme JW#1. The labeled toxin produced by incorporation of [9,10(-3)H]palmitate induced phytotoxic symptoms identical to unlabeled material, indicating it had full biological activity. The area of necrosis on treated leaves was similar in light and dark treated plants. Using liquid scintillation counting to quantify radioactivity in excised plant parts, over 95% of the [3H]FB1 radioactivity applied to leaves of light or dark-treated plants was recovered from the treated leaf. When [3H]FB1 was applied to a wound site on target plants, severe damage occurred at the site of FB1 application and in tissue above the site. These results indicate that FB1 applied to intact surfaces of target plants exhibits primarily contact activity. Translocation of FB1 is limited, occurring only when FB1 is applied to a wound site, and it results in damage to tissue above the point of application, indicating that FB1 is xylem mobile.


The outcome of the vertical vein left intact during the surgery for total anomalous venous connection and its effects on ventricular functions.
We examined the fate of intact vertical vein during long-term follow-up and its effects on ventricular functions. Eighty one patients with all types of total anomalous pulmonary venous connection (TAPVC) were operated. Thirty-one patients with supracardiac type TAPVC were examined in our retrospective cohort study. Groups were evaluated with respect to left ventricle area, volume, end-systolic, end diastolic diameter, early and late term mortality and properties of pulmonary hypertensive crisis. Vertical vein was left open in 14 patients and it was ligated in 17 patients. Wilcoxon rank and Mann- Whitney U tests were used to compare variables between groups. After mean follow up of 48±36 months, vertical vein closed spontaneously in 3 patients. Two of them were closed surgically due to elevated shunt flow and there was spontaneous closure in one patient who had the highest pulmonary artery pressure amongst others postoperatively. Preoperative left ventricular area, volume, end-systolic diameter and end-diastolic diameter values of the patients increased from 3.5± 0.9 mm(2)-2.9±0.9 mm(3)-14±5 mm-8±4.5 mm to 8±3.3 mm(2)-16±8.7 mm(3)-27±6.7 mm-17±4.8 mm at the postoperative period in Group 1, and from 6.8±3.5 mm(2)-8.4±8.7 mm(3)-15±6.4 mm-9±5.3 mm to 7.4±5.2 mm(2)-16±1.7 mm(3)-21.7±5 mm-13.5±4.1 mm in Group 2, respectively (p=0.02, p=0.039, p=0.054, and p=0.07, respectively). Vertical vein remains intact in most of the patients and may be closed spontaneously in patients with elevated pulmonary resistance. Intact vertical vein both decompresses the left ventricular cavity in patients with decreased left ventricular compliance until the ventricular muscle adapts to new workload and improves ventricular functions on long term follow up. Therefore, we conclude that vertical vein should be left open in selected patients.


Is job strain a major source of cardiovascular disease risk?
Empirical studies on job strain and cardiovascular disease (CVD), their internal validity, and the likely direction of biases were examined. The 17 longitudinal studies had the highest validity ratings. In all but two, biases towards the null dominated. Eight, including several of the largest, showed significant positive results; three had positive, nonsignificant findings. Six of nine case-control studies had significant positive findings; recall bias leading to overestimation appears to be fairly minimal. Four of eight cross-sectional studies had significant positive results. Men showed strong, consistent evidence of an association between exposure to job strain and CVD. The data of the women were more sparse and less consistent, but, as for the men, most of the studies probably underestimated existing effects. Other elements of causal inference, particularly biological plausibility, corroborated that job strain is a major CVD risk factor. Additional intervention studies are needed to examine the impact of ameliorating job strain upon CVD-related outcomes.


Azacarbazole n-3 and n-6 polyunsaturated fatty acids ethyl esters nanoemulsion with enhanced efficacy against Plasmodium falciparum.
Alternative therapies are necessary for the treatment of malaria due to emerging drug resistance. However, many promising antimalarial compounds have poor water solubility and suffer from the lack of suitable delivery systems, which seriously limits their activity. To address this problem, we synthesized a series of azacarbazoles that were evaluated for antimalarial activity against D10 (chloroquine-sensitive) and W2 (chloroquine-resistant) strains of P. falciparum. The most active compound, 9H-3-azacarbazole (3), was encapsulated in a novel o/w nanoemulsion consisting of ethyl esters of polyunsaturated fatty acids n-3 and n-6 obtained from flax oil as the oil phase, Smix (Tween 80 and Transcutol HP) and water. This formulation was further analyzed using transmission electron microscopy, dynamic light scattering and in vitro and in vivo studies. It was shown that droplets of the 3-loaded nanosystem were spherical, with satisfactory stability, without cytotoxicity towards fibroblasts and intestinal cell lines at concentrations corresponding to twice the IC50 for P. falciparum. Moreover, the nanoemulsion with this type of oil phase was internalized by Caco-2 cells. Additionally, pharmacokinetics demonstrated rapid absorption of compound 3 (tmax = 5.0 min) after intragastric administration of 3-encapsulated nanoemulsion at a dose of 0.02 mg/kg in mice, with penetration of compound 3 to deep compartments. The 3-encapsulated nanoemulsion was found to be 2.8 and 4.2 times more effective in inhibiting the D10 and W2 strains of the parasite, respectively, compared to non-encapsulated 3. Our findings support a role for novel o/w nanoemulsions as delivery vehicles for antimalarial drugs.


An Explanation for Neptune's Ring Arcs.
The Voyager mission revealed a complex system of rings and ring arcs around Neptune and uncovered six new satellites, four of which occupy orbits well inside the ring region. Analysis of Voyager data shows that a radial distortion with an amplitude of approximately 30 kilometers is traveling through the ring arcs, a perturbation attributable to the nearby satellite Galatea. Moreover, the arcs appear to be azimuthally confined by a resonant interaction with the same satellite, yielding a maximum spread in ring particle semimajor axes of 0.6 kilometer and a spread in forced eccentricities large enough to explain the arcs' 15-kilometer radial widths. Additional ring arcs discovered in the course of this study give further support to this model.


Salting-out assisted extraction method coupled with hydrophilic interaction liquid chromatography for determination of selected β-blockers and their metabolites in human urine.
In this study, a new analytical method was developed and validated for the simultaneous analysis of β-blockers (metoprolol, propranolol, carvedilol) and their metabolites (5'-hydroxycarvedilol, O-desmethylcarvedilol, α-hydroxymetoprolol, O-desmethylmetoprolol, 5-hydroxypropranolol) in human urine. A salting-out assisted liquid-liquid extraction (SALLE) procedure was used for sample preparation. Several parameters affecting the extraction efficiency and method sensitivity including the type and volume of the extraction solvent, the type and quantity of the inorganic salt, extraction time and sample pH were investigated. Hydrophilic interaction liquid chromatography-ultraviolet detection (HILIC-UV) was used for the determination of all analytes. During method development, the effects of mobile phase components (type, pH, concentration of salt, organic modifier type and content, flow rate, column temperature) on the retention and separation of β-blockers and metabolites on the five different HILIC columns were examined. The method was linear for concentrations ranging from 0.1 to 8.0μg/mL, with determination coefficients higher than 0.993 for all analytes. The limits of quantification were in the range from 0.1 to 0.2μg/mL. Intra- and inter-day precision ranged from 0.1 to 8.9%, and accuracy was within±13% interval for all analytes. Under the optimized conditions, extraction efficiency was greater than 83.4% for determined compounds. The validated method was then applied to the measurement of β-blockers and their metabolites in human urine samples.


Desmoglein-2 interaction is crucial for cardiomyocyte cohesion and function.
We determined the contribution of the desmosomal cadherin desmoglein-2 to cell-cell cohesion in cardiomyocytes. In the intercalated disc, providing mechanical strength and electrical communication between adjacent cardiomyocytes, desmoglein-2 is closely associated with N-cadherin and gap junctions. We studied intercalated discs of HL-1 cardiomyocytes by immunostaining of desmoglein-2 and N-cadherin. Cohesion was measured using a liberase-based dissociation-assay and compared with cell-free single-molecule atomic force microscopy measurements. L-tryptophan caused irregular desmoglein-2 condensation, weakened cell-cell cohesion and impaired both homophilic desmoglein-2 and N-cadherin trans-interaction, whereas l-phenylalanine had no effect. L-tryptophan did not affect N-cadherin localization and its inhibitory effect on cell-cohesion and desmoglein-2 binding, but not on N-cadherin interaction, was blocked by a desmoglein-specific tandem peptide. Moreover, Ca(2+)-depletion, desmoglein-2 knockdown, a desmoglein-specific single peptide and certain desmoglein-2 mutations associated with arrhythmogenic cardiomyopathy reduced cell-cell cohesion, whereas cell adhesion was strengthened by desmoglein-2 overexpression. Since single peptide did not interfere with N-cadherin trans-interaction, these data indicate that (i) desmoglein-2 binding is crucial for cardiomyocyte cohesion and (ii) L-tryptophan reduced both desmoglein-2 and N-cadherin binding, whereas single and tandem peptide can be used to specifically target desmoglein-2-mediated adhesion. L-tryptophan and single peptide also induced ultrastructural alterations of areae compositae. Functional analyses at the organ level revealed reduced cardiomyocyte function and inefficient response to adrenergic stimulation in both L-tryptophan- and single peptide-challenged murine Langendorff hearts paralleled by redistribution of connexin 43 in L-tryptophan-treated heart slices. Our data demonstrate that desmoglein-2 plays a critical role in cardiomyocyte cohesion and function.


Incidence of neutropenia in patients with ticlopidine/Ginkgo biloba extract combination drug for vascular events: A post-marketing cohort study.
The ticlopidine/Ginkgo biloba ext. combination drug (Yuclid) is used as an antiplatelet agent for prevention of vascular events since its approval in 2008. The purpose of this study is to explore the safety of ticlopidine/Ginkgo biloba combination, mainly regarding the incidence of neutropenia, through a post-marketing surveillance study. From March 2009 to October 2015, a total of 4839 subjects had been enrolled in this study. The enrollments were conducted by 152 doctors of 89 hospitals according to the regulations for post-marketing surveillance programs in Korea. If a subject was administered the drug once, he/she was included in the safety analysis set for any adverse events and bleedings, and the primary safety evaluation regarding neutropenia was conducted in subjects who completed 3-month blood test follow-up. We predefined that 1% reduction in neutropenia incidence by ticlopidine/Ginkgo biloba ext. combination from the previously reported incidence of ticlopidine of 2.3% was clinically meaningful. Among the safety analysis set of 4831 patients (99.8% of the enrolled subjects), 3150 (65.1%) completed evaluation for neutropenia at 3 months which is the primary safety endpoint. The major causes of dropout were no follow-up visit at 3 months (n = 1016) and violation of the follow-up period (n = 503). Nine patients experienced neutropenia (Absolute neutrophil count [ANC] ≤ 1200mm3) and the estimated cumulative incidence at 3 months is 0.29% (95% confidence interval, 0.13%- 0.54%). Severe neutropenia (ANC ≤ 450mm3) did not occur in any patients. The incidence of neutropenia with addition of Ginkgo biloba ext. to ticlopidine may be lower than the previously reported incidence of neutropenia with ticlopidine, which needs to be confirmed in randomized controlled trials.


Persistent prelimbic cortex activity contributes to enhanced learned fear expression in females.
Anxiety disorders, such as post-traumatic stress, are more prevalent in women and are characterized by impaired inhibition of learned fear and medial prefrontal cortex (mPFC) dysfunction. Here we examined sex differences in fear extinction and mPFC activity in rats. Females showed more learned fear expression during extinction and its recall, but not fear conditioning. They also showed more spontaneous fear recovery and more contextual fear before extinction and its recall. Moreover, enhanced learned fear expression in females was associated with sustained prelimbic (PL) cortex activity. These results suggest that sex differences in learned fear expression may involve persistent PL activation.


Healthy store programs and the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), but not the Supplemental Nutrition Assistance Program (SNAP), are associated with corner store healthfulness.
In response to lack of access to healthy foods, many low-income communities are instituting local healthy corner store programs. Some stores also participate in the United States Department of Agriculture's Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) and the Supplemental Nutrition Assistance Program (SNAP). This study used two assessment tools to compare the healthfulness of offerings at stores participating in local healthy store programs (upgraded stores), WIC, and/or SNAP to that of similar non-participating stores. Based on store audits conducted in 315 New Jersey corner stores in 2014, we calculated healthy food availability scores using subsections of the Nutrition Environment Measures Survey for Corner Stores (NEMS-CS-Availability) and a short-form corner store audit tool (SCAT). We used multivariable regression to examine associations between program participation and scores on both instruments. Adjusting for store and block group characteristics, stores participating in a local healthy store program had significantly higher SCAT scores than did non-participating stores (upgraded: M = 3.18, 95% CI 2.65-3.71; non-upgraded: M = 2.52, 95% CI 2.32-2.73); scores on the NEMS-CS-Availability did not differ (upgraded: M = 12.8, 95% CI 11.6-14.1; non-upgraded: M = 12.5, 95% CI 12.0-13.0). WIC-participating stores had significantly higher scores compared to non-participating stores on both tools. Stores participating in SNAP only (and not in WIC) scored significantly lower on both instruments compared to non-SNAP stores. WIC-participating and non-SNAP corner stores had higher healthfulness scores on both assessment tools. Upgraded stores had higher healthfulness scores compared to non-upgraded stores on the SCAT.


Effect of reduced oxygen tension on chondrogenesis and osteogenesis in adipose-derived mesenchymal cells.
Recent studies have demonstrated that adipose-derived mesenchymal cells (AMCs) offer great promise for cell-based therapies because of their ability to differentiate toward bone, cartilage, and fat. Given that cartilage is an avascular tissue and that mesenchymal cells experience hypoxia during prechondrogenic condensation in endochondral ossification, the goal of this study was to understand the influence of oxygen tension on AMC differentiation into bone and cartilage. In vitro chondrogenesis was induced using a three-dimensional micromass culture model supplemented with TGF-beta1. Collagen II production and extracellular matrix proteoglycans were assessed with immunohistochemistry and Alcian blue staining, respectively. Strikingly, micromasses differentiated in reduced oxygen tension (2% O(2)) showed markedly decreased chondrogenesis. Osteogenesis was induced using osteogenic medium supplemented with retinoic acid or vitamin D and was assessed with alkaline phosphatase activity and mineralization. AMCs differentiated in both 21 and 2% O(2) environments. However, osteogenesis was severely diminished in a low-oxygen environment. These data demonstrated that hypoxia strongly inhibits in vitro chondrogenesis and osteogenesis in AMCs.


Impact of maternal postnatal depression on cognitive development of young children.
Ninety four women and their first born children took part in a longitudinal study of maternal mental health during pregnancy and after delivery. The children's cognitive functioning was assessed at age 4 using the McCarthy scales, without knowledge of the mothers' psychiatric history or current health. As expected girls performed slightly better than boys and children from middle class and professional families did better than children from working class homes, as did children whose mothers had achieved at least one A level at school. Significant intellectual deficits were found in the children whose mothers had suffered with depression, but only when this depression occurred in the first year of the child's life. Marital conflict and a history of paternal psychiatric problems were independently linked with lower cognitive test scores; together with a working class home background these were the only factors that contributed to the deleterious effect of maternal postnatal depression.


SEARCH international case-control study of childhood brain tumours: role of index pregnancy and birth, and mother's reproductive history.
A series of co-ordinated population-based case-control studies of childhood brain tumours (CBT) was undertaken under the auspices of the Surveillance of Environmental Aspects Related to Cancer in Humans (SEARCH) programme of the International Agency for Research on Cancer (IARC) to evaluate, inter alia, the risk in relation to characteristics of the index pregnancy and birth, and maternal reproductive history. Subjects comprised 1218 cases aged 0-19 years and 2223 controls. Risk estimates were calculated by unconditional logistic regression, adjusted for age, sex, centre and mother's years of schooling, for all types of CBT combined as well as for four groups defined by histopathology (astrologlial tumours, primitive neuroectodermal tumours of the brain, 'other glial' tumours and 'other histological types') and for five age groups (0-1, 0-4, 5-9, 10-14, 15-19 years). Use of anaesthetic 'gas' was associated with an increased risk of CBT (OR = 1.5, 95% CI [1.1, 2.0]), apparent in children aged 0-4 years (OR = 2.4, 95% CI [1.4, 4.1]) and for astroglial tumours (OR = 1.6, 95% CI [1.1, 2.2]) with non-significantly increased relative risks for each of the other histological groups. However, not all centre-specific relative risks were elevated. No other aspect of the index pregnancy, delivery and early neonatal period or of the mother's previous reproductive history was associated with risk for CBT.


Prospective ten-month exercise intervention in premenarcheal girls: positive effects on bone and lean mass.
Enhancement of bone mineral acquisition during growth may be a useful preventive strategy against osteoporosis. The aim of this study was to explore the lean mass, strength, and bone mineral response to a 10-month, high-impact, strength-building exercise program in 71 premenarcheal girls, aged 9-10 years. Lean body mass, total body (TB), lumbar spine (LS), proximal femur (PF), and femoral neck (FN) bone mineral were measured using the Hologic QDR 2000+ bone densitometer. Strength was assessed using a grip dynamometer and the Cybex isokinetic dynamometer (Cybex II). At baseline, no significant difference in body composition, pubertal development, calcium intake, physical activity, strength, or bone mineral existed between groups. At completion, there were again no difference in height, total body mass, pubertal development, calcium intake, or external physical activity. In contrast, the exercise group gained significantly more lean mass, less body fat content, greater shoulder, knee and grip strength, and greater TB, LS, PF, and FN BMD (exercise: TB 3.5%, LS 4.8%, PF 4.5%, and FN 12.0%) compared with the controls (controls: TB 1.2%, LS 1.2%, PF 1.3%, and FN 1.7%). TB bone mineral content (BMC), LS BMC, PF BMC, FN BMC, LS bone mineral apparent density (BMAD), and FN bone area also increased at a significantly greater rate in the exercise group compared with the controls. In multiple regression analysis, change in lean mass was the primary determinant of TB, FN, PF, and LS BMD accrual. Although a large proportion of bone mineral accrual in the premenarcheal skeleton was related to growth, an osteogenic effect was associated with exercise. These results suggest that high-impact, strength building exercise is beneficial for premenarcheal strength, lean mass gains, and bone mineral acquisition.


IMPACT OF KRAS MUTATIONS IN CLINICAL FEATURES IN COLORECTAL CANCER.
KRAS mutations are important events in colorectal carcinogenesis, as well as negative predictors of response to EGFR inhibitors treatment. To investigate the association of clinical-pathological features with KRAS mutations in colorectal cancer patients treated. Data from 69 patients with colorectal cancer either metastatic at diagnosis or later, were retrospectively analyzed. The direct sequencing and pyrosequencing techniques were related to KRAS exon 2. The mutation diagnosis and its type were determined. KRAS mutation was identified in 43.4% of patients. The most common was c.35G>T (p.G12V), c.35G>A (p.G12D) and c.38G>A (p.G13D). No correlation was found between KRAS mutation and age (p=0.646) or gender (p=0.815). However, mutated group had higher CEA levels at admission (p=0.048) and codon 13 mutation was associated with involvement of more than one metastatic site in disease progression (p=0.029). Although there was no association between primary tumor site and mutation diagnosis (p=0.568), primary colon was associated with worse overall survival (p=0.009). The KRAS mutation was identified in almost half of patients. Mutated KRAS group had higher levels of CEA at admission and the mutation at codon 13 was associated with involvement of more than one metastatic site in the course of the disease. Colon disease was associated with the worst overall survival.


Effect of GH/IGF-1 on Bone Metabolism and Osteoporsosis.
Background. Growth hormone (GH) and insulin-like growth factor (IGF-1) are fundamental in skeletal growth during puberty and bone health throughout life. GH increases tissue formation by acting directly and indirectly on target cells; IGF-1 is a critical mediator of bone growth. Clinical studies reporting the use of GH and IGF-1 in osteoporosis and fracture healing are outlined. Methods. A Pubmed search revealed 39 clinical studies reporting the effects of GH and IGF-1 administration on bone metabolism in osteopenic and osteoporotic human subjects and on bone healing in operated patients with normal GH secretion. Eighteen clinical studies considered the effect with GH treatment, fourteen studies reported the clinical effects with IGF-1 administration, and seven related to the GH/IGF-1 effect on bone healing. Results. Both GH and IGF-1 administration significantly increased bone resorption and bone formation in the most studies. GH/IGF-1 administration in patients with hip or tibial fractures resulted in increased bone healing, rapid clinical improvements. Some conflicting results were evidenced. Conclusions. GH and IGF-1 therapy has a significant anabolic effect. GH administration for the treatment of osteoporosis and bone fractures may greatly improve clinical outcome. GH interacts with sex steroids in the anabolic process. GH resistance process is considered.


A cross-sectional study of the presence of United Kingdom (UK) plastic surgeons on social media.
To determine the uptake and usage of websites and social media (SM) by UK consultant (attending) plastic surgeons. Professional profiles of full BAPRAS members were searched on Facebook, Twitter, LinkedIn, RealSelf, YouTube, ResearchGate in May 2013. Additional surgeons were identified from the follower lists of @BAPRASvoice and @BAAPSMedia. Website ownership was determined on Google. Searches were repeated three times. Dual BAAPS-BAPRAS members were identified from www.baaps.org.uk. There were 156 (48.3%) dual BAAPS-BAPRAS members and 36 BAPRAS-only members. Fifty seven (18%) surgeons had no account on any platform whereas 266 (82%) were on at least one platform. One hundred and sixty four (51%) had personal websites whilst 37 (11%) had profiles on partnership websites. One hundred and sixteen (36%) had no website presence whilst 2% had websites under construction. The platform most surgeons use is LinkedIn (52%) whilst smaller proportions used Facebook (4%) and Twitter (22%). Surgeons had a mean of 126 (range: 0-3270) Twitter followers and 368 (range: 7-3786) fans/'likes' of their Facebook profiles. Time spent in postgraduate practice was not predictive of website ownership or SM use. However, dual BAAPS-BAPRAS members were significantly more likely to own a personal website, Twitter, RealSelf and YouTube accounts. There has been an increase in the uptake of social media by UK plastic surgeons, especially in those with aesthetic surgery interests. However, very few surgeons have optimised their web presence. Continued education and appropriate usage guidance may promote uptake, particularly by reconstructive surgeons.


Functional expression of carnitine/organic cation transporter OCTN1/SLC22A4 in mouse small intestine and liver.
Carnitine/organic cation transporter (OCTN1/SLC22A4) accepts various therapeutic agents as substrates in vitro and is expressed ubiquitously, although its function in most organs has not yet been examined. The purpose of the present study was to evaluate functional expression of OCTN1 in small intestine and liver, using octn1 gene knockout [octn1(-/-)] mice. After oral administration of [(3)H]ergothioneine ([(3)H]ERGO), a typical substrate of OCTN1, the amount of [(3)H]ERGO remaining in the small intestinal lumen was much higher in octn1(-/-) mice than in wild-type mice. In addition, uptake of [(3)H]ERGO by human embryonic kidney 293 cells heterologously expressing OCTN1 gene product and uptake of [(3)H]ERGO at the apical surface of intestinal everted sacs from wild-type mice were inhibited by OCTN1 substrates, tetraethylammonium and verapamil. Immunohistochemical analysis revealed that OCTN1 is localized on the apical surface of small intestine in mice and humans. These results suggest that OCTN1 is responsible for small intestinal absorption of [(3)H]ERGO. However, the plasma concentration of [(3)H]ERGO after oral administration was higher in octn1(-/-) mice than in wild-type mice, despite the lower absorption in octn1(-/-) mice. This was probably because of efficient hepatic uptake of [(3)H]ERGO, as revealed by integration plot analysis; the uptake clearance was close to the hepatic plasma flow rate. The uptake of [(3)H]ERGO by isolated hepatocytes was minimal, whereas [(3)H]ERGO uptake was observed in isolated nonparenchymal cells. This finding is consistent with immunostaining of OCTN1 in liver sinusoids. Thus, our results indicate that OCTN1 is functionally expressed in nonparenchymal liver cells.


Prevalence of obsessive-compulsive disorder in the British nationwide survey of child mental health.
Obsessive-compulsive disorder (OCD) is a disorder that appears to be under-diagnosed and under-treated, despite the evidence for effective treatments. There are variable estimates of OCD prevalence in the under-16s and published rates give little indication of age trends. The aim of this study was to establish the prevalence and associates of OCD in young people aged 5-15 years. The method was a nationwide (UK) epidemiological study of rates of psychiatric disorder in 5-15 year olds (1999 British Child Mental Health Survey): 10,438 children were assessed. Twenty-five children with OCD were identified (weighted overall prevalence 0.25%; 95% CI 0.14-0.35), with prevalence rising exponentially with increasing age. Compared with normal controls, children with OCD were more likely to be from lower socio-economic class and of lower intelligence. Only three of these children had been seen by specialist children's services. Although OCD is rare in young children, the rate increases towards the adult rates at puberty. Children with OCD have additional psychosocial disadvantage. The majority of the childhood cases identified in this survey appear to have been undetected and untreated.


Thermal conductivity of graphene and graphite: collective excitations and mean free paths.
We characterize the thermal conductivity of graphite, monolayer graphene, graphane, fluorographane, and bilayer graphene, solving exactly the Boltzmann transport equation for phonons, with phonon-phonon collision rates obtained from density functional perturbation theory. For graphite, the results are found to be in excellent agreement with experiments; notably, the thermal conductivity is 1 order of magnitude larger than what found by solving the Boltzmann equation in the single mode approximation, commonly used to describe heat transport. For graphene, we point out that a meaningful value of intrinsic thermal conductivity at room temperature can be obtained only for sample sizes of the order of 1 mm, something not considered previously. This unusual requirement is because collective phonon excitations, and not single phonons, are the main heat carriers in these materials; these excitations are characterized by mean free paths of the order of hundreds of micrometers. As a result, even Fourier's law becomes questionable in typical sample sizes, because its statistical nature makes it applicable only in the thermodynamic limit to systems larger than a few mean free paths. Finally, we discuss the effects of isotopic disorder, strain, and chemical functionalization on thermal performance. Only chemical functionalization is found to play an important role, decreasing the conductivity by a factor of 2 in hydrogenated graphene, and by 1 order of magnitude in fluorogenated graphene.


Prebiotic synthesis of 2-deoxy-d-ribose from interstellar building blocks promoted by amino esters or amino nitriles.
Understanding the prebiotic genesis of 2-deoxy-d-ribose, which forms the backbone of DNA, is of crucial importance to unravelling the origins of life, yet remains open to debate. Here we demonstrate that 20 mol% of proteinogenic amino esters promote the selective formation of 2-deoxy-d-ribose over 2-deoxy-d-threopentose in combined yields of ≥4%. We also demonstrate the first aldol reaction promoted by prebiotically-relevant proteinogenic amino nitriles (20 mol%) for the enantioselective synthesis of d-glyceraldehyde with 6% ee, and its subsequent conversion into 2-deoxy-d-ribose in yields of ≥ 5%. Finally, we explore the combination of these two steps in a one-pot process using 20 mol% of an amino ester or amino nitrile promoter. It is hence demonstrated that three interstellar starting materials, when mixed together with an appropriate promoter, can directly lead to the formation of a mixture of higher carbohydrates, including 2-deoxy-d-ribose.


Dynamics of enhancer chromatin signatures mark the transition from pluripotency to cell specification during embryogenesis.
The generation of distinctive cell types that form different tissues and organs requires precise, temporal and spatial control of gene expression. This depends on specific cis-regulatory elements distributed in the noncoding DNA surrounding their target genes. Studies performed on mammalian embryonic stem cells and Drosophila embryos suggest that active enhancers form part of a defined chromatin landscape marked by histone H3 lysine 4 mono-methylation (H3K4me1) and histone H3 lysine 27 acetylation (H3K27ac). Nevertheless, little is known about the dynamics and the potential roles of these marks during vertebrate embryogenesis. Here, we provide genomic maps of H3K4me1/me3 and H3K27ac at four developmental time-points of zebrafish embryogenesis and analyze embryonic enhancer activity. We find that (1) changes in H3K27ac enrichment at enhancers accompany the shift from pluripotency to tissue-specific gene expression, (2) in early embryos, the peaks of H3K27ac enrichment are bound by pluripotent factors such as Nanog, and (3) the degree of evolutionary conservation is higher for enhancers that become marked by H3K27ac at the end of gastrulation, suggesting their implication in the establishment of the most conserved (phylotypic) transcriptome that is known to occur later at the pharyngula stage.


Paclitaxel combined with harmine inhibits the migration and invasion of gastric cancer cells through downregulation of cyclooxygenase-2 expression.
Cyclooxygenase-2 (COX-2) has a critical role in the invasiveness and metastasis of gastric cancer. In addition, paclitaxel (PTX) and harmine (HM) were reported to be potential therapeutic drug candidates for cancer therapy; however, the synergistic antitumor effect of PTX and HM combined treatment on the human gastric cancer cells remains to be elucidated. The aim of the present study was to evaluate the effects of PTX and/or HM on the cell migration and invasion in two human gastric cancer cell lines, SGC-7901 and MKN-45. MTT assay was used to detect the growth inhibition induced by PTX and HM. The Transwell assay was employed to assess the effects of PTX and HM on the cell migration and invasion. The expression levels of COX-2 and matrix metalloproteinase-9 (MMP-9) were analyzed by western blot analysis. The results demonstrated that PTX and HM inhibited cell proliferation in a dose-dependent manner. Individually PTX and HM were able to inhibit the migration and invasion of two human gastric cancer cells; however, the combination of PTX and HM exerted synergistic effects on migration and invasion inhibition, with downregulation of COX-2 and matrix metalloproteinase (MMP)-9. In conclusion, the results of the present study indicated that combination chemotherapy using PTX with HM exerted an antitumor effect, which may be implicated for the treatment of gastric cancer. Of note, the combination of the two drugs inhibited migration and invasion more effectively compared with each drug alone, the mechanism of which proceeded via the downregulation of COX-2 expression.


Inclusion of quebracho tannin extract in a high-roughage cattle diet alters digestibility, nitrogen balance, and energy partitioning.
Condensed tannins (CT) might improve animal and system-level efficiency due to enhanced protein efficiency and reduced CH4. This study evaluated the impact of quebracho tannin (QT) extract fed at 0%, 1.5%, 3%, and 4.5% of dry matter (DM), within a roughage-based diet on apparent digestibility of DM, organic matter (OM), fibrous fractions, and N retention and energy partitioning of growing steers (236 ± 16 kg BW). A Latin rectangle design with eight animals and four periods was used to determine the whole-animal exchange of CO2, O2, and CH4 as well as the collection of total feces and urine over a 48-h period, using two open-circuit, indirect calorimetry respiration chambers. Following the removal of steers from respiration chambers, rumen inoculum was collected to determine ruminal parameter, including volatile fatty acids (VFA) and ammonia. Animals were fed a 56.5% roughage diet at 1.7% BW (dry matter basis). Dry matter and gross energy intakes were influenced by the level of QT inclusion (P ≤ 0.036). Digestibility of DM, OM, and N was reduced with QT inclusion (P < 0.001), and fiber digestibility was slightly impacted (P > 0.123). QTs altered the N excretion route, average fecal N-to-total N ratio excreted increased 14%, and fecal N-to-urinary N ratio increased 38% (P < 0.001) without altering the retained N. Increased fecal energy with QT provision resulted in reduced dietary digestible energy (DE) concentration (Mcal/kg DM; P = 0.024). There were no differences in urinary energy (P = 0.491), but CH4 energy decreased drastically (P = 0.007) as QT inclusion increased. Total ruminal VFA concentration did not differ across treatments, but VFA concentration increased linearly with QT inclusion (P = 0.049). Metabolizable energy (ME) was not affected by the QT rate, and the conversion efficiency of DE-to-ME did not differ. Heat energy decreased (P = 0.013) with increased QT provision likely due to changes in the DE intake, but there was no difference in retained energy. There were no differences for retained energy or N per CO2 equivalent emission produced (P = 0.774 and 0.962, respectively), but improved efficiency for energy retention occurred for 3% QT. We concluded that QT provided up to 4.5% of dry matter intake (about 3.51% of CT, dry matter basis) does not affect N and energy retention within the current setting. Feeding QT reduced energy losses in the form of CH4 and heat, but the route of energy loss appears to be influenced by the rate of QT inclusion.


Advanced tracer techniques to monitor synaptic activity.
The two approaches presented here bypass postsynaptic receptors as indicators of quantal release, and thus they can provide information which is clearly distinct from that obtained with standard electrophysiological techniques. Indeed, the inherently variable responsiveness of the postsynaptic membrane makes it an unreliable indicator of presynaptic activity and this has fueled a lot of controversy, particularly in the area of synaptic plasticity. A major advantage of these two methods is their ability to detect changes at the single bouton level. This offers a lot of advantages including the possibility to study the functional role for exo-endocytosis but also plasticity against a background of great variability among a large number of synapses. The spatial resolving power of FM1-43 and anti-synaptotagmin antibodies may be valuable in future studies of spread of LTP between neighboring synapses and in the mapping the pattern of neuronal activity in complex networks of neurons.


Differential Recruitment of Parietal Cortex during Spatial and Non-spatial Reach Planning.
The planning of goal-directed arm reaching movements is associated with activity in the dorsal parieto-frontal cortex, within which multiple regions subserve the integration of arm- and target-related sensory signals to encode a motor goal. Surprisingly, many of these regions show sustained activity during reach preparation even when target location is not specified, i.e., when a motor goal cannot be unambiguously formed. The functional role of these non-spatial preparatory signals remains unresolved. Here this process was investigated in humans by comparing reach preparatory activity in the presence or absence of information regarding upcoming target location. In order to isolate the processes specific to reaching and to control for visuospatial attentional factors, the reaching task was contrasted to a finger movement task. Functional MRI and electroencephalography (EEG) were used to characterize the spatio-temporal pattern of reach-related activity in the parieto-frontal cortex. Reach planning with advance knowledge of target location induced robust blood oxygenated level dependent and EEG responses across parietal and premotor regions contralateral to the reaching arm. In contrast, reach preparation without knowledge of target location was associated with a significant BOLD response bilaterally in the parietal cortex. Furthermore, EEG alpha- and beta-band activity was restricted to parietal scalp sites, the magnitude of the latter being correlated with reach reaction times. These results suggest an intermediate stage of sensorimotor transformations in bilateral parietal cortex when target location is not specified.


Quality of life and its determinants among community re-integrated soldiers with permanent disabilities following traumatic limb injuries.
To assess health-related-quality of life (HRQOL) after the end of war and its associated factors among soldiers with permanent limb disabilities in Sri Lanka METHODS: A cross-sectional study was conducted among 500 rehabilitated and socially re-integrated Army soldiers having permanent disability following traumatic limb injuries during the last year of ethnic conflict of Sri Lanka. Females, soldiers with no updated data, and soldiers having permanent deafness/total blindness/total brain or spinal cord injury were excluded. The sample was randomly selected from updated 'Disabled Category Registries'. Personal and injury data were collected through postal questionnaire. HRQOL was assessed using SF-36 tool validated for Sri Lankan soldiers. SF-36 scores obtained on HRQOL related to overall physical health (mean = 57.1; SD = 19) and mental health (mean = 58.8; SD = 17.2) were comparable. HRQOL was best in social functioning (mean = 73.4; SD = 24) and worst in vitality (mean = 48.4; SD = 15.5). Factors significantly associated with better HRQOL related to the overall physical and mental health were aged over 35 years, being married, being assigned duties, having only upper/lower limbs affected and absence of co-disability. Having duties assigned demonstrated better QOL in almost all domains. None were associated with income or education. Physical and mental health components contribute equally to the HRQOL of rehabilitated soldiers with permanent limb disabilty. Their poorer HRQOL was determined by certain person- and injury-related factors. These need to be addressed during re-integration and post-rehabilitation services.


Revision surgery for cervical spondylotic myelopathy: surgical results and outcome.
The role of additional or revision surgery in patients with cervical spondylotic myelopathy (CSM) is challenging. Postoperative pseudoarthrosis, instability, hardware failure, and recurrent cervical stenosis are conditions that require detailed clinical and radiographic assessment to define the pathology and assess the need for surgical decompression and fusion. The purpose of this study is to assess the neurological outcome, radiological outcome, and complications of patients undergoing additional or revision surgery for CSM. Between 2002 and 2006, 30 patients with CSM and postoperative pseudoarthrosis, instability, hardware failure, or recurrent stenosis underwent surgical decompression and stabilization. The specific procedure was selected according to each patient's medical condition, cervical sagittal alignment, and extent of stenosis. All patients underwent an anterior, posterior, or combined anterior and posterior decompression and instrumented fusion. The charts of these patients were reviewed to assess neurological and radiographic outcomes. Twenty-five patients (83%) improved postoperatively as measured by the Nurick Myelopathy Scale over a mean follow-up period of 19 months (range, 2-64 mo). The overall complication rate was 27%, consisting of transient monoradiculopathy (7%), dysphagia (10%), and infection (7%). The incidence of nonunion during the follow-up period was 3%. Although patients with CSM and postoperative pseudoarthrosis, instability, hardware failure, or junctional stenosis who require revision surgery may risk a substantial likelihood of surgical complications (25% in this series), a significant proportion of patients may experience improved neurological outcomes. In our experience, the cervical sagittal alignment and the extent of stenosis are critical factors to consider when selecting the eventual procedure.


Oligomerization of bacterially expressed H1N1 recombinant hemagglutinin contributes to protection against viral challenge.
Vaccination is the most effective intervention to prevent influenza and control the spread of the virus. Alternatives are needed to the traditional egg-based vaccine strategy for a more rapid response to new outbreaks. Two different hemagglutinin (HA) fragments (rHA11-326 and rHA153-269) derived from influenza A virus subtype H1N1 were expressed in Escherichia coli and characterized by immunoblot, gel filtration, hemagglutination, and competitive binding assays. rHA11-326 included neutralizing epitopes and the trimerization domain, whereas rHA153-269 included only the head of HA with the neutralizing epitopes. Mice were immunized with rHA11-326 or rHA153-269, and sera were tested for the presence of neutralizing antibodies. Mice were then challenged with H1N1 and infection severity was monitored. rHA11-326 trimerized, whereas rHA153-269 was unable to form oligomers. Both rHA11-326 and rHA153-269 elicited the production of neutralizing antibodies, but only oligomerized rHA11-326 protected against live virus challenges in mice. This study demonstrated that bacterially expressed HA was capable of folding properly and eliciting the production of neutralizing antibodies, and that HA oligomerization contributed to protection against viral challenge. Therefore, prokaryotic-derived vaccine platforms can provide antigenic and structural requirements for viral protection, as well as allow for the rapid and cost-effective incorporation of multiple antigens for broader protection.


First Report of Neofusicoccum parvum Associated with Grapevine Trunk Diseases in Croatia.
In September 2010, during survey of diseased grapevines (Vitis vinifera L.) in vineyards at localities Zmajevac (BZ), Orahovica (SO), Cilipi (KC), and Novalja (PN), symptoms characteristic of grapevine trunk diseases (GTD) (3) were observed, showing on cross-sectioned cordons and trunks as brown, wedge-shaped perennial cankers and/or dark streaking of the wood. In Croatia, these symptoms were traditionally associated with Eutypa Tul. & C.Tul. and with fungi from Diaporthaceae (2). From affected grapevines (cvs. Grasevina, Pinot bijeli, Malvazija dubrovacka, and Gegic), samples of symptomatic cordons and trunks were collected (n ≥ 35). To isolate the causal agents from the samples, woodchips of symptomatic tissue, surface-sterilized in 2% sodium hypochlorite for 2 min, were placed on potato dextrose agar amended with streptomycin sulphate (50 μg/ml) and incubated for 7 days at 25°C in darkness. A percentage of samples (72, 15, 27, and 54% from BZ, SO, KC, and PN, respectively) yielded fungal colonies with abundant aerial mycelium, initially white, but turning olivaceous grey after 5 days. From these colonies, monohyphal isolates were obtained and pycnidial formation stimulated by cultivation on 2% water agar with stems of plant species Foeniculum vulgare Mill. at 25°C under diffuse light for 3 weeks. Pycnidia contained conidia that were hyaline, unicellular, ellipsoid with round apices and truncated bases, and thin walled with smooth surface. Dimensions of conidia (n ≥ 50) were (12.8) 15.3 ± 1.4 (17.6) × (5.4) 6.3 ± 0.8 (7.6) μm, with length/width ratio (2.0) 2.5 ± 0.5 (3.2). Based on morphological data, species Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips was suspected (1). For molecular identification, isolates BZ330, SO334, KC342, and PN121 were used for PCR to amplify internal transcribed spacer region and partial translation elongation factor 1-alpha gene, using primers ITS5/ITS4 and EF1-728F/EF1-986R, respectively. Obtained sequences were shown to be identical between the four isolates (GenBank: KF296318, KF296319) and when compared with sequences for reference N. parvum isolate CMW9080 (AY236942, AY236887) they showed >99% homology, confirming the isolates as species N. parvum. Pathogenicity tests were done by inoculation of detached green shoots (GS) and lignified canes (LC) (n = 5) of grapevine cv. Skrlet by either mycelial plugs of the same four isolates, or sterile agar plugs for the controls. Inoculated GS were kept in flasks with sterile water in a glasshouse for 10 days, and LC in humid dark chambers for 30 days, at 25°C. Resulting vascular necrosis measured 62 to 81 mm (GS) and 215 to 246 mm (LC), but was absent on controls. Koch's postulates were satisfied by successful reisolation of N. parvum only from plants inoculated with mycelial plugs. N. parvum has been recognized as a serious grapevine pathogen, causing similar symptoms worldwide (3). To our knowledge, this is the first report of N. parvum associated with GTD in Croatia, and due to its relatively high incidence at surveyed localities, it could present considerable threat, particularly for neighboring vine growing regions. Diplodia seriata De Not., a weak pathogen (3), was also identified from a percentage of samples in this survey. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) J. Kaliterna et al. Arh. Hig. Rada Toksikol. 63:471, 2012. (3) J. R. Urbez-Torres. Phytopathol. Mediterr. 50(Suppl.):S5, 2011.


Vitamin C in the treatment of acquired immune deficiency syndrome (AIDS).
My previous experience with the utilization of ascorbic acid in the treatment of viral diseases led me to hypothesize that ascorbate would be of value in the treatment of AIDS (acquired immune deficiency syndrome). Preliminary clinical evidence is that massive doses of ascorbate (50-200 grams per 24 hours) can suppress the symptoms of the disease and can markedly reduce the tendency for secondary infections. In combination with usual treatments for the secondary infections, large doses of ascorbate will often produce a clinical remission which shows every evidence of being prolonged if treatment is continued. This clinical remission is achieved despite continuing laboratory evidence of helper T-cell suppression. There may be a complete or partial destruction of the helper T-cells during an initial infection that does not necessitate a continuing toxicity from some source to maintain a permanent or prolonged helper T-cell suppression. However, it is possible ascorbate may prevent that destruction if used adequately during that prodrome period. Emphasis is put upon the recognition and treatment of the frequent intestinal parasites. Food and chemical sensitivities occur frequently in the AID syndrome and may aggravate symptoms considered to be part of the AID syndrome. A topical C-paste has been found very effective in the treatment of herpes simplex and, to a lesser extent, in the treatment of some Kaposi's lesions. Increasingly, clinical research on other methods of treating AIDS is being "contaminated" by patients taking ascorbate.


Mitochondrial lineages in Ladin-speaking communities of the eastern Alps.
European mitochondrial alleles cluster into five haplogroups. Haplogroup 2 is rare in general, but represents more than half of the few known sequences among Ladin speakers of the Alps. Here we describe DNA diversity in control region I of the hypervariable D-loop in 43 Ladins, and in 25 Italian speakers. Analysis of these data, and of previously published sequences, confirms a high degree of differentiation among Ladins and their geographical neighbours. This cannot be regarded as a simple effect of isolating factors, geographic or linguistic, as diversity is high within Ladin communities too. Rather, allele genealogies, population trees, and principal component analysis suggest a relationship between Ladin and Near Eastern samples. Two evolutionary hypotheses seem compatible with these findings. The view whereby Ladins could be descended from Palaeolithic inhabitants of the Alps is supported by the identification, in this study, of the probable ancestral haplotype of group 2, never previously observed in central Europe. Alternatively, a comparatively recent, Neolithic immigration of the ancestors of current Ladin speakers seems consistent with recent linguistic theories. In both cases, the number of lineages present, and their extensive diversity, are not compatible with a serious bottleneck in the Ladin population's history.


The gut microbiota modulates host amino acid and glutathione metabolism in mice.
The gut microbiota has been proposed as an environmental factor that promotes the progression of metabolic diseases. Here, we investigated how the gut microbiota modulates the global metabolic differences in duodenum, jejunum, ileum, colon, liver, and two white adipose tissue depots obtained from conventionally raised (CONV-R) and germ-free (GF) mice using gene expression data and tissue-specific genome-scale metabolic models (GEMs). We created a generic mouse metabolic reaction (MMR) GEM, reconstructed 28 tissue-specific GEMs based on proteomics data, and manually curated GEMs for small intestine, colon, liver, and adipose tissues. We used these functional models to determine the global metabolic differences between CONV-R and GF mice. Based on gene expression data, we found that the gut microbiota affects the host amino acid (AA) metabolism, which leads to modifications in glutathione metabolism. To validate our predictions, we measured the level of AAs and N-acetylated AAs in the hepatic portal vein of CONV-R and GF mice. Finally, we simulated the metabolic differences between the small intestine of the CONV-R and GF mice accounting for the content of the diet and relative gene expression differences. Our analyses revealed that the gut microbiota influences host amino acid and glutathione metabolism in mice.


Adjunctive β2-agonist treatment reduces glycogen independently of receptor-mediated acid α-glucosidase uptake in the limb muscles of mice with Pompe disease.
Enzyme or gene replacement therapy with acid α-glucosidase (GAA) has achieved only partial efficacy in Pompe disease. We evaluated the effect of adjunctive clenbuterol treatment on cation-independent mannose-6-phosphate receptor (CI-MPR)-mediated uptake and intracellular trafficking of GAA during muscle-specific GAA expression with an adeno-associated virus (AAV) vector in GAA-knockout (KO) mice. Clenbuterol, which increases expression of CI-MPR in muscle, was administered with the AAV vector. This combination therapy increased latency during rotarod and wirehang testing at 12 wk, in comparison with vector alone. The mean urinary glucose tetrasaccharide (Glc4), a urinary biomarker, was lower in GAA-KO mice following combination therapy, compared with vector alone. Similarly, glycogen content was lower in cardiac and skeletal muscle following 12 wk of combination therapy in heart, quadriceps, diaphragm, and soleus, compared with vector alone. These data suggested that clenbuterol treatment enhanced trafficking of GAA to lysosomes, given that GAA was expressed within myofibers. The integral role of CI-MPR was demonstrated by the lack of effectiveness from clenbuterol in GAA-KO mice that lacked CI-MPR in muscle, where it failed to reverse the high glycogen content of the heart and diaphragm or impaired wirehang performance. However, the glycogen content of skeletal muscle was reduced by the addition of clenbuterol in the absence of CI-MPR, as was lysosomal vacuolation, which correlated with increased AKT signaling. In summary, β2-agonist treatment enhanced CI-MPR-mediated uptake and trafficking of GAA in mice with Pompe disease, and a similarly enhanced benefit might be expected in other lysosomal storage disorders.


Population genetic structure, genetic diversity, and natural history of the South American species of Nothofagus subgenus Lophozonia (Nothofagaceae) inferred from nuclear microsatellite data.
The effect of glaciation on the levels and patterns of genetic variation has been well studied in the Northern Hemisphere. However, although glaciation has undoubtedly shaped the genetic structure of plants in the Southern Hemisphere, fewer studies have characterized the effect, and almost none of them using microsatellites. Particularly, complex patterns of genetic structure might be expected in areas such as the Andes, where both latitudinal and altitudinal glacial advance and retreat have molded modern plant communities. We therefore studied the population genetics of three closely related, hybridizing species of Nothofagus (N. obliqua, N. alpina, and N. glauca, all of subgenus Lophozonia; Nothofagaceae) from Chile. To estimate population genetic parameters and infer the influence of the last ice age on the spatial and genetic distribution of these species, we examined and analyzed genetic variability at seven polymorphic microsatellite DNA loci in 640 individuals from 40 populations covering most of the ranges of these species in Chile. Populations showed no significant inbreeding and exhibited relatively high levels of genetic diversity (H E = 0.502-0.662) and slight, but significant, genetic structure (R ST = 8.7-16.0%). However, in N. obliqua, the small amount of genetic structure was spatially organized into three well-defined latitudinal groups. Our data may also suggest some introgression of N. alpina genes into N. obliqua in the northern populations. These results allowed us to reconstruct the influence of the last ice age on the genetic structure of these species, suggesting several centers of genetic diversity for N. obliqua and N. alpina, in agreement with the multiple refugia hypothesis.


An examination of the cross-sectional geometrical properties of the long bone diaphyses of Holocene foragers from Roonka, South Australia.
This study examines long bone diaphyseal rigidity and shape of hunter-gatherers at Roonka to make inferences about subsistence strategies and mobility of inhabitants of semi-arid southeastern Australia. Roonka is a cemetery site adjacent to the Lower Murray River, which contains over 200 individuals buried throughout the Holocene. Archaeological evidence indicates that populations living near this river corridor employed mobile, risk averse foraging strategies. This prediction of lifestyle was tested by comparing the cross-sectional geometric properties of the humerus, radius, ulna, femur, tibia, and fibula of individuals from Roonka to samples of varying subsistence strategies. Bilateral asymmetry of the upper limb bones was also examined. Roonka males and females have moderately high lower limb diaphyseal rigidity and shape. In the upper limb, females have low rigidity and bilateral asymmetry while males have moderately high rigidity and bilateral asymmetry. This pattern is similar to other foraging groups from Australia and southern Africa that have behaviorally adapted to arid and semi-arid environments. Lower limb results suggest that populations in the Lower Murray River Valley had relatively elevated foraging mobility. Upper limb rigidity and bilateral asymmetry indicate a sexual division of labor at Roonka. Females resemble other samples that had mixed subsistence strategies that involved hunting, gathering, and processing tasks. Males display a pattern similar to groups that preferentially hunted large game, but that supplemented this source with smaller game and riverine resources.


Exercise test and glucose homeostasis in rats treated with alloxan during the neonatal period or fed a high calorie diet.
Animal models appear well-suited for studies into the role of exercise in the prevention of non-insulin-dependent diabetes mellitus (NIDDM). The aim of the present study was to analyze glucose homeostasis and blood lactate during an exercise swimming test in rats treated with alloxan during the neonatal period and/or fed a high calorie diet from weaning onwards. Rats were injected with alloxan (200 mg/kg, i.p.) or vehicle (citrate buffer) at 6 days of age. After weaning, rats were divided into four groups and fed either a balanced diet or a high-caloric diet as follows: C, control group (vehicle + normal diet); A, alloxan-treated rats fed the normal diet; H, vehicle-treated rats fed the high-caloric diet; and HA, alloxan-treated rats fed the high-caloric diet. Fasting serum glucose levels were higher in groups A and AH compared with the control group. The Homeostatic Model Assessment index varied in the groups as follows: H>A>HA = C. There were no differences in free fatty acids or blood lactate concentrations during the swim test. Alloxan-treated rats fed a normal or high-caloric diet have the potential to be used in studies analyzing the role physical exercise plays in the prevention of NIDDM.


Playing 'hide-and-seek' with factor H: game-theoretical analysis of a single nucleotide polymorphism.
As a part of the complement system, factor H regulates phagocytosis and helps differentiate between a body's own and foreign cells. Owing to mimicry efforts, some pathogenic microorganisms such as Candida albicans are able to bind factor H on their cell surfaces and, thus, become similar to host cells. This implies that the decision between self and foreign is not clear-cut, which leads to a classification problem for the immune system. Here, two different alleles determining the binding affinity of factor H are relevant. Those alleles differ in the SNP Y402H; they are known to be associated with susceptibility to certain diseases. Interestingly, the fraction of both alleles differs in ethnic groups. The game-theoretical model proposed in this article explains the coexistence of both alleles by a battle of the sexes game and investigates the trade-off between pathogen detection and protection of host cells. Further, we discuss the ethnicity-dependent frequencies of the alleles. Moreover, the model elucidates the mimicry efforts by pathogenic microorganisms.


Effect of apple polyphenol concentrate on lipid metabolism in rats under experimental insulin resistance.
Obesity is strongly associated with an increased risk of developing insulin resistance as the metabolic indicator of prediabetes and a major risk factor in diabetes mellitus type 2 pathogenesis. Medicinal products obtained from apples can be used as potent prophylactic and therapeutic remedies in treatment of diabetes mellitus. Experiment was designed to study the effect of total apple polyphenol food concentrate on lipid metabolism under experimental IR. Male Wistar rats weighting 180-210 g were used in the experiment. IR was induced by high-calorie diet enriched with fructose. The effect of total apple polyphenol food concentrate was compared with the action of epigallocatechin gallate and quercetin. To estimate the alterations in lipid metabolism in liver homogenate were measured triacylglycerols, free fatty acids, total phospholipids, TBA-reactive substance and conjugated dienes contents. In blood serum were measured total lipids, triacylglycerols, cholesterol, total phospholipids and reduced glutathione levels. The obtained results indicated that feeding rats with high-calorie diet enriched with fructose caused the dyslipidemia and oxidative stress development. The administration of quercetin, epigallocatechin gallate and total apple polyphenol food concentrate improved disorders of lipid metabolism and pro-oxidant-antioxidant homeostasis. Total apple polyphenol food concentrate had a more pronounced effect on studied indices that is probably due to synergism and additive effect of extract numerous components.


The Origins and Maintenance of Female Genital Modification across Africa : Bayesian Phylogenetic Modeling of Cultural Evolution under the Influence of Selection.
We present formal evolutionary models for the origins and persistence of the practice of Female Genital Modification (FGMo). We then test the implications of these models using normative cross-cultural data on FGMo in Africa and Bayesian phylogenetic methods that explicitly model adaptive evolution. Empirical evidence provides some support for the findings of our evolutionary models that the de novo origins of the FGMo practice should be associated with social stratification, and that social stratification should place selective pressures on the adoption of FGMo; these results, however, are tempered by the finding that FGMo has arisen in many cultures that have no social stratification, and that forces operating orthogonally to stratification appear to play a more important role in the cross-cultural distribution of FGMo. To explain these cases, one must consider cultural evolutionary explanations in conjunction with behavioral ecological ones. We conclude with a discussion of the implications of our study for policies designed to end the practice of FGMo.